Sulfatase-activated fluorophores for rapid discrimination of mycobacterial species and strains

Abstract

Most current diagnostic tests for tuberculosis do not reveal the species or strain of pathogen causing pulmonary infection, which can lead to inappropriate treatment regimens and the spread of disease. Here, we report an assay for mycobacterial strain assignment based on genetically conserved mycobacterial sulfatases. We developed a sulfatase-activated probe, 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)-sulfate, that detects enzyme activity in native protein gels, allowing the rapid detection of sulfatases in mycobacterial lysates. This assay revealed that mycobacterial strains have distinct sulfatase fingerprints that can be used to judge both the species and lineage. Our results demonstrate the potential of enzyme-activated probes for rapid pathogen discrimination for infectious diseases.Mycobacterium tuberculosis | chemical biology | fluorescence | enzyme assay | hydrolase

A single diamagnetic catalyCEST MRI contrast agent that detects cathepsin B enzyme activity by using a ratio of two CEST signals

“…Other peptide sequences can be incorporated into the synthesis scheme to create catalyCEST MRI contrast agents that sense other proteases (26). Other classes of enzymes that modify an aryl functional group may also be detected using this platform technology, such as esterases (7), phosphatases (27), and sulfatases (28). The salicylic acid moiety may be replaced by other molecules that have intramolecular hydrogen bonding that causes highly shifted CEST signals and are less likely to undergo slow degradation (11,17,29).…”

An Expanded Set of Fluorogenic Sulfatase Activity Probes

“…[6] We recently reported on the use of fluorogenic probes to examine mycobacterial sulfatase activity in a variety of species and strains. [7] In that work, we developed a new fluorogenic probe, 7-hydroxy-9H-(1,3-dichloro-9,9-dimethylacridin-2-one)(DDAO)-sulfate, which was used to detect sulfatase activity in protein gel-resolved mycobacterial lysates. The assay revealed that mycobacteria have distinct sulfatase activity patterns, or “fingerprints”.…”

Synthesis and Evaluation of Cytosolic Phospholipase A2 Activatable Fluorophores for Cancer Imaging

“…DDAO has also been conjugated to form activatable fluorophores through conjugation of its hydroxyl group resulting in photoactivatable and sulfatase activatable probes. 42, 43 …”